Microfl uidic Blood Cell Sorting: Now and Beyondme-web.engin.umich.edu/ibbl/pdf/2013_Small_Yu.pdf · One main use of blood in medicine is for blood transfu-sion, in which components
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Zeta Tak For Yu , Koh Meng Aw Yong , and Jianping Fu *
Blood plays an important role in homeostatic regulation with each of its cellular components having important therapeutic and diagnostic uses. Therefore, separation and sorting of blood cells hasa been of a great interest to clinicians and researchers. However, while conventional methods of processing blood have been successful in generating relatively pure fractions, they are time consuming, labor intensive, and are not optimal for processing small volume blood samples. In recent years, microfl uidics has garnered great interest from clinicians and researchers as a powerful technology for separating blood into different cell fractions. As microfl uidics involves fl uid manipulation at the microscale level, it has the potential for achieving high-resolution separation and sorting of blood cells down to a single-cell level, with an added benefi t of integrating physical and biological methods for blood cell separation and analysis on the same single chip platform. This paper will fi rst review the conventional methods of processing and sorting blood cells, followed by a discussion on how microfl uidics is emerging as an effi cient tool to rapidly change the fi eld of blood cell sorting for blood-based therapeutic and diagnostic applications.
cells such as CTCs from patient blood [ 9 ] and (iv) targeting
particular WBC subpopulations at various status.
Conventional blood cell sorting methods include the
use of antibodies and their specifi city to protein markers
on blood cells of interest. Antibody-based approaches pos-
sess the advantage of high specifi city and sensitivity but are
limited by the quality and high cost of antibodies. Label-free
separation of cellular components of blood by their physical
properties such as cell density and size has also been widely
used; however, high-resolution separation of blood cells is dif-
fi cult with centrifugation or size-based fi ltration approaches
using fi brous membranes or track-etched polycarbonate fi l-
ters. In addition, conventional macroscale blood cell isolation
methods require a large volume of blood and involve many
manual interventions prone to introducing artifacts, and they
also commonly require skilled technicians and well-equipped,
expensive laboratories.
Novel microfl uidics and lab-on-a-chip (LOC) tech-
nology developments have been gaining in importance in
recent years as effi cient and powerful approaches for high-
throughput blood cell separation, owing to their precise
control of fl uid behavior and the ability to scale down the
required sample volume and achieve continuous non-inva-
sive molecular and functional analysis of blood cells down
to the single-cell level. In addition, exploring unique fl uidic
transport phenomena in confi ning microfl uidic environments
and integrating both physical and biochemical methods and
analytical assays in a single-chip format provide comprehen-
sive capabilities of integrated microfl uidic approaches for
blood cell sorting and analysis over conventional macroscale
methods.
Over the last decade, there have been many novel
developments in designing highly integrated and functional
microfl uidic devices and systems that incorporate different
approaches for the separation and sorting of blood cells as
well as rare cells in blood such as CTCs. Some of these micro-
fl uidic cell separation and sorting techniques have been dis-
cussed in several recent informative and insightful reviews. [ 10 ]
Particularly, the blood-on-a-chip review [ 10g ] by Toner and
Irimia has discussed thoroughly the complexity and asso-
ciated challenges of handling and processing blood using
emerging microfl uidics technologies. However, there are
Dr. Z. T. F. Yu,[+] Dr. K. M. Aw Yong,[+] Prof. J. Fu Integrated Biosystems and Biomechanics Laboratory University of Michigan Ann Arbor, Michigan , USA E-mail: [email protected]
Dr. Z. T. F. Yu, Dr. K. M. Aw Yong, Prof. J. Fu Department of Mechanical Engineering University of Michigan Ann Arbor, Michigan , USA
Prof. J. Fu Department of Biomedical Engineering University of Michigan Ann Arbor, Michigan , USA[+]These authors contributed equally to this work.
2D acoustic concentration – 4999 0.025 to 0.1 RBCs 0.15 to 0.2 >94 – [100]
a) Ratio of volumes for samples diluted in diluent before chip loading; b) Ratio of amounts of target components before and after chip processes; c) Ratio of amounts of target components compared
devices integrated with PDMS microfi ltration membranes
had successfully achieved on-chip isolation, enrichment,
and functional analysis of PBMCs as well as subpopulations
of WBCs such as CD14+ monocytes from lysed and whole
blood specimens with high fl ow rate (up to 20 mL min −1 ) and
excellent cell purity (>97%). [ 11,39 ] Another material that has
been successfully utilized for microfabrication of fi ltration
membranes was Parylene-C, a biocompatible polymer that
between the pillars of 5 µm) only trapped cancer cells but not
blood cells. The isolation effi ciency was reported to be >80%
for breast and colon cancer cell lines spiked in blood samples
under a low pressure operation.
Although a precise control of the microfl uidic fi lter
geometry is necessary for achieving optimal cell sorting
performance, improved operations of these microfl uidic fi l-
ters can also result in increased cell separation and sorting
effi ciency. Such an attempt was successfully demonstrated
using a structural ratchet mechanism created using funnel-
shaped microscale constrictions under an oscillatory fl ow,
Figure 2 B. [ 38 ] When a cell suspension containing mouse
lymphoma cells (MLCs) and human peripheral blood
mononuclear cells (PBMCs) was injected through a 2D fi lter
array consisted of funnel constrictions, PBMCs that were
smaller and more deformable could easily fl ow across the
constrictions in forward fl ow while larger and less deform-
able MLCs were excluded. When a reversal fl ow was
applied, MLCs that were trapped at the constrictions would
be released to unclog the funnel constrictions. PBMCs how-
ever were not able to pass back through the constrictions
since the cells had to overcome a smaller opening on the lee-
ward side of the funnel constrictions. By repeating the for-
ward and reversal fl ow, MLCs and PBMCs were successfully
separated with enhanced separation effi ciencies and purities.
Figure 2. Physical fi ltration. (A) One-dimensional (1D) fi lter arrays. Left: Linear fi lter array. Reproduced with permission. [ 32 ] Copyright 1994, American Association for Clinical Chemistry. Middle: Slanted cross-fl ow fi lter array. Reproduced with permission. [ 33 ] Copyright 2010, Elsevier. Right: Diffusive cross-fl ow fi lter array. Reproduced with permission. [ 34 ] Copyright 2006, Royal Society of Chemistry. (B) Two-dimensional (2D) fi lter arrays. Left: Gradient array with successively narrower fi lters along the fl ow direction. Reproduced with permission. [ 35 ] Copyright 2004, The Institute of Electrical and Electronics Engineers. Middle: Crescent-shaped pillar arrays. Reproduced with permission. [ 37 ] Copyright 2009, Springer. Right: Ratchet pillars with reversal fl ow. Reproduced with permission. [ 38 ] Copyright 2012, Royal Society of Chemistry. (C) 2D microfi ltration membranes made of PDMS. Reproduced with permission. [ 11 ] Copyright 2013, Wiley-VCH.
hemodynamic phenomena have been the focus of attempts
by researchers to design microfl uidic networks that mimic in
vivo fl ow conditions to enhance sorting and separation of dif-
ferent blood components.
Inertial focusing, for example, has attracted much atten-
tion to the microfl uidic community in recent years as a novel
strategy to control and steer particles in microfl uidic chan-
nels. [ 46 ] As its name suggests, inertial focusing makes use of
inertial forces generated as a result of fl uidic fl ow within a
confi ning microchannel. Shear-gradient lift and wall-induced
lift generate a net lift force that drives particles toward equi-
librium positions within the microchannel cross-section,
turning an initial homogeneous microparticle stream into
a highly focused microparticle stream within a short dis-
tance, Figure 3 A. [ 47 ] The inertial force is mainly regulated
by two parameters: the Dean number and the ratio of par-
possesses good mechanical strength and
fl exibility. [ 40 ] The Parylene microfi ltration
membranes (with the membrane surface
area up to 36 mm 2 and porosity about 7%
– 15%) could contain well-defi ned pores of
different geometries (circular, oval-shaped,
and rectangular pores) with critical dimen-
sions down to a few microns. The Parylene
microfi ltration membranes were success-
fully applied for capturing CTCs from
1 mL of whole blood in less than 5 min,
achieving 90% capture effi ciency, 90%
cell viability, and 200-fold sample enrich-
ment. [ 40 ] It is worth noting that other
microfi ltration membranes made by nickel
electroplating [ 41 ] or through direct uses of
commercial fi ltration papers [ 42 ] or plastic
sheets [ 43 ] have also been successfully
incorporated into microfl uidic blood cell
sorting devices.
The fi ltrate purity achieved by physical
fi ltration of blood cells is inevitably com-
promised by stowaway cells. Blood cells
are fairly deformable and can easily pass
through a slit or constriction smaller than
the cell size. Furthermore, there is a signifi -
cant size overlap among various blood cell
subpopulations as well as CTCs. In order
to enhance separation resolution and thus
purity for size-based blood cell separation,
our research laboratory and others have
recently developed a strategy to combine
microfi ltration membranes with antibody-
conjugated microbeads for isolation and
enrichment of subpopulations of WBCs
as well CTCs. Microbeads conjugated with
antibodies could bind target WBCs [ 11,39 ]
or CTCs [ 44 ] to increase the apparent size
difference between target cells and other
blood cells, thus enhancing separation
resolution and purity from subsequent
cell fi ltration process using microfi ltration
membranes, Figure 2 C. Another recent
study further utilized instability of fl uid fl ow (such as the
Taylor–Gortler instability phenomenon) to enhance mixing
of antibody-coated microbeads and blood cells in microfl u-
idic channels to improve the bead-cell conjugation process
before downstream cell fi ltration process to isolate and
enrich CTCs. [ 45 ]
3.2. Hydrodynamic Mechanisms and Hemodynamic Phenomena
The human circulatory system is made up of a complex net-
work of blood vessels interacting as microfl uidic systems for
transporting blood. The rich and complex interactions among
blood cells, blood plasma and confi ning blood vessels at
the microscale level have allowed establishments of unique
blood fl ow characteristics. Blood related hydrodynamic and
Figure 3. Hydrodynamic mechanisms. (A) Inertial focusing. Left: In serpentine channel. Reproduced with permission. [ 47 ] Copyright 2007, The National Academy of Sciences, U.S.A. Middle: In high aspect-ratio channel. Reproduced with permission. [ 48 ] Copyright 2011, Royal Society of Chemistry. Right: In curvilinear channel. Reproduced with permission. [ 49 ] Copyright 2013, Nature Publishing Group. (B) Hydrodynamic focusing. Top: Bilateral sheath streams. Reproduced with permission. [ 54a ] Copyright 2006, American Chemical Society. Bottom: A single-sided sheath stream. Reproduced with permission. [ 12a ] Copyright 2010, John Wiley & Sons. (C) Hydrophoretic focusing. Reproduced with permission. [ 55a ] Copyright 2011, Royal Society of Chemistry. (D) Deterministic lateral displacement (DLD). Left: Separation of blood. Reproduced with permission. [ 57 ] Copyright 2006, The National Academy of Sciences, U.S.A. Middle: Sorting of RBCs. Reproduced with permission. [ 58 ] Copyright 2012, Royal Society of Chemistry. Right: I-shaped pillar array. Reproduced with permission. [ 12c ] Copyright 2013, Nature Publishing Group.
Another unique hemodynamic behavior is the Zweifach-
Fung effect, also known as the bifurcation law, which
describes the tendency of RBCs to travel into a vessel with
a higher fl ow rate as it encounters a bifurcating region. This
effect was successfully applied to separate plasma when
blood fl owed through a microchannel with multiple high-
fl ow-resistance branches (Figure 4 ). [ 66 ]
Microfl uidic sorting of blood cells using physical fi ltra-
tion and hydrodynamic and hemodynamic mechanisms may
unavoidably introduce stresses to cells, and such unintended
stimulations may alter molecular expressions and even cel-
lular phenotypes of blood cells. [ 67 ] A recent relevant study
compared activation of polymorphonuclear leukocytes
(PMNs) when undergoing centrifugation in conventional
centrifuges and curvilinear microchannels. The percentage of
activated PMNs in both systems was found low compared to
treatments such as RBC lysis. [ 50 ] In conclusion, it is important
to conduct comparative assays to evaluate the effect of phys-
ical manipulations (such as physical fi ltration and shear fl ow)
of cells in microfl uidic environment on blood cell behaviors.
These control assays will also be valuable for determining the
optimized microfl uidic device geometries and operational
parameters.
Figure 4. Hemodynamic phenomena. Top left: Fahraeus-Lindqvist effect. Reproduced with permission. [ 63 ] Copyright 2011, Royal Society of Chemistry. Bottom left: Cell-free effect and plasma skimming effect. Reproduced with permission. [ 64 ] Copyright 2006, IOS Press. Middle: Cell-free effect. Reproduced with permission. [ 65 ] Copyright 2005, American Chemical Society. Right: Zweifach-Fung effect. Reproduced with permission. [ 66 ] Copyright 2006, Royal Society of Chemistry.
printed antibody array was able to capture specifi c T-cells
from RBC-depleted human whole blood for multiplexed
detection of cytokines secreted from T-cells. Such multi-para-
metric analyses of T-cell functions are valuable for diagnosis
and monitoring drug response of infectious diseases such as
AIDS and TB.
Our research laboratory has recently utilized the dif-
ferential adhesion preferences of cancer cells to nanorough
glass surfaces as compared to intrinsically non-adherent
blood cells to achieve effi cient CTC capture without using
capture antibodies. [ 13b ] To this end, we developed a simple yet
precise controlled method to generate random nanorough-
ness on glass surfaces using reactive ion etching (RIE). These
nanoroughened glass surfaces were shown to effi ciently cap-
ture different kinds of cancer cells derived from different tis-
sues (i.e., MCF-7, MDA-MB-231, HeLa, PCS, and SUM-149)
spiked in blood samples, Figure 5 D. [ 13b ]
Affi nity-based techniques for sorting blood cells can be
very specifi c and thus cell purity resulted from such methods
are superior to those achieved by other microfl uidic cell
sorting methods. However, it remains a challenge to release
captured cells from an antibody coated surface without using
enzymes or shear stress. To allow releasing target cells after
diagnostics (Figure 5 A). [ 74 ] When leukemia cell lines with
different P-selectin affi nities fl owed over the ridges, a lateral
driving force induced by hydrophoresis enabled differential
displacements along the ridge direction, thus enabling sepa-
ration of leukemia cells in the device. Another more recent
microfl uidic cell sorting device utilized a plain feature-free
surface functionalized with P-selectin ligands in a slanted
strip pattern to generate lateral forces, in order to separate
WBCs from RBCs in blood samples (Figure 5 B). [ 13a ] Simi-
larly, such spatial patterning of adhesive ligands was applied
successfully for separation of WBCs and CTCs from blood
samples [ 75 ] as well as to create quadruplex capture of multiple
WBC types. [ 76 ]
Glycosylation, the addition of sugar moieties to macro-
molecules such as proteins or lipids occurs frequently in
nature and affects cell adhesion as well as protein structure
and function. [ 77 ] In diseases such as cancer, aberrant glyco-
sylation or the dysregulation of lectins (proteins that bind
sugars) can occur, and both have been of great interest to
researchers as potential targets for personalized cancer
therapy. [ 78 ] It is conceivable that such a phenomenon can
be utilized within a microfl uidic system as an alternative
to antibody-based approaches for cell capture and sorting.
One such study involved functionalizing a multivalent sur-
face with galactose and using its affi nity for binding its
partner galectin-3 (overexpressed on the surface of meta-
static cells) to capture cancer cells. [ 79 ] The use of carbohy-
drates rather than antibodies also possesses the potential
of allowing for the continuous monitoring of cancer cell
mutations of particular antigenic structures. [ 79b ] In addi-
tion, these sugar-functionalized surfaces allow the study
Figure 5. Surface affi nity and topography. (A) Affi nity based micro/nanostructures. Left: Antibody-coated pillar array. Reproduced with permission. [ 69 ] Copyright 2007, Nature Publishing Group. Middle: antibody-coated nanopillars. Reproduced with permission. [ 71 ] Copyright 2011, Wiley-VCH. Right: Ligand-receptor interaction. Reproduced with permission. [ 74 ] Copyright 2012, Royal Society of Chemistry. (B) Array by surface patterning. Reproduced with permission. [ 13a ] Copyright 2013, Nature Publishing Group. (C) Array by robotic printing. Reproduced with permission. [ 80 ] Copyright 2008, Royal Society of Chemistry. (D) Nanotopography. Reproduced with permission. [ 13b ] Copyright 2013, American Chemical Society. (E) Capture and release of cells. Reproduced with permission. [ 13c ] Copyright 2012, Wiley-VCH.
linker to bind antibodies to solid surfaces, selective capture
and release of target cells via photochemical cleavage was
demonstrated. [ 83 ]
From a biological point of view, all affi nity-based methods
for capture and enrichment of blood cells will introduce
some specifi c interactions between antibodies or ligands
and cell surface antigens. While some surface antigens may
function simply as structural constituents of cell membranes,
others may be involved in important signaling pathways that
regulate molecular and cellular functions. Hence, there are
concerns whether these affi nity-based methods will alter the
physiology of cells after cell sorting and isolation processes.
Figure 6. Magnetophoresis. (A) Magnetic cell separation. Left: Trapping by cavities. Reproduced with permission. [ 84a ] Copyright 2012, Royal Society of Chemistry. Right: Positive and negative selections. Reproduced with permission. [ 12b ] Copyright 2013, American Association for the Advancement of Science. (B) Self-assembled magnetic bead array. Reproduced with permission. [ 86 ] Copyright 2010, The National Academy of Sciences, U.S.A. (C) Inherent paramagnetic properties of cells. Reproduced with permission. [ 87 ] Copyright 2006, Institution of Engineering and Technology.
different blood cells for separation while the second cat-
egory makes use of biological differences, such as surface
protein markers to help discriminate between different cell
types. While highly effective, there are limitations to such
macroscale approaches, such as long processing time, high
cost and the availability of good antibodies, and the amount
of blood sample required. Owing to precise control over
the cell microenvironment and the ability to scale down the
operation to very small volumes of blood, recent advances in
microfl uidics have been gaining in importance as effi cient and
powerful approaches for high-throughput blood cell sorting
and separation as well as non-invasive molecular and func-
tional analysis down to a single-cell resolution. The fi eld of
blood cell sorting using microfl uidics keeps evolving, and we
foresee that new trends will lie in the following categories:
(i) integration of multiple cell separation modules for advanced
sorting of target blood cells, (ii) integration of upstream
microfl uidic blood cell sorting with downstream molecular, cel-
lular and functional analysis on the single-chip platform, and
(iii) complex, highly integrated microfl uidic devices and sys-
tems with novel functionalities for high-throughput, high-con-
tent blood analysis. However, we need to reckon that current
microfl uidic cell sorting technologies are still facing signifi cant
challenges that need to be fully addressed in the near future,
in order to fulfi ll their true potential and realize their impact
on blood-based therapeutic and diagnostic applications. These
challenges include the laborious fabrication process required
for generating defect-free, intricate 3D microfl uidic networks
and their associated complex control and operation, and
interfaces between delicate, highly integrated microfl uidic
systems with conventional macroscopic instruments. None-
theless, given the potential and signifi cant advantages of well-
controlled microfl uidic environment and the rich mechanisms
that are available for manipulation of blood cells, we envision
that microfl uidic blood sorting systems may one day become a
mainstay in the clinical and research laboratories.
a cell, electrophysiological properties and molecular and cel-
lular phenotypes of the cell may also be altered.
Acoustophoresis is another contact-free, label-free
method that can focus particles laterally within microchan-
nels. The generation of an ultrasound radiation force in an
acoustically soft medium confi ned within acoustically rigid
microchannel walls requires two criteria: (i) channel dimen-
sions in concert with ultrasound frequencies, and (ii) a
channel dimension that matches an integral multiple of half-
wavelength in suspending fl uid. [ 97 ] Under ultrasound fi elds,
particles can move to either the pressure node or anti node
of the ultrasound fi eld in the microfl uidic channel. As the
magnitude of the acoustic force depends on particle size, den-
sity and compressibility, acoustophoresis has been utilized
to isolate blood components such as peripheral blood pro-
genitor cells (PBPCs) for transplantation, [ 98 ] blood plasma
for biomarker detection, [ 99 ] and CTCs for cancer diagnostics
(Figure 7 B). [ 97 ] Moreover, 200-fold enrichment of RBCs was
achieved under 2D acoustic standing wave in a microfl u-
idic setting (Figure 7 B). [ 100 ] It is worth noting that a recent
study systematically examined the potential adverse effect
of microfl uidic acoustophoresis on phenotypic changes of
microglia cells, human prostate cancer cells, human throm-
bocytes and WBCs, including their viability and proliferation
and infl ammatory responses. This study suggested that ultra-
sonic actuation with the operating voltage less than 10 Vpp
would have negligible adverse impacts on many important
cellular functions. [ 101 ]
4. Concluding Remarks
Conventional methods of sorting and separation of blood
cells can be classifi ed into two main categories and can
be used alone or together. The fi rst category falls under
making use of the differences in physical properties between
Figure 7. Electrical methods and acoustophoresis. (A) Left: Electrowetting-on-dielectric (EWOD). Reproduced with permission. [ 91 ] Copyright 2010, American Institute of Physics. Right: Dielectrophoresis (DEP). Reproduced with permission. [ 93 ] Copyright 2012, Wiley-VCH. (B) Left: 1D cell alignment. Reproduced with permission. [ 97 ] Copyright 2012, American Chemical Society. Right: 2D cell concentration. Reproduced with permission. [ 100 ] Copyright 2012, Royal Society of Chemistry.
15 ( 2 ), 149 – 157 ; e) H. W. Hou , A. A. S. Bhagat , W. C. Lee , S. Huang , J. Han , C. T. Lim , Micromachines-Basel 2011 , 2 ( 3 ), 319 – 343 ; f) A. Lenshof , T. Laurell , Chem. Soc. Rev. 2010 , 39 ( 3 ), 1203 – 1217 ; g) M. Toner , D. Irimia , Annu. Rev. Biomed. Eng. 2005 , 7 , 77 – 103 ; h) L. Yu , S. R. Ng , Y. Xu , H. Dong , Y. J. Wang , C. M. Li , Lab Chip 2013 , 13 ( 16 ), 3163 – 3182 .
[11] W. Chen , N. T. Huang , B. Oh , R. H. Lam , R. Fan , T. T. Cornell , T. P. Shanley , K. Kurabayashi , J. Fu , Adv. Healthc. Mater. 2013 .
[12] a) M. Kim , S. Mo Jung , K. H. Lee , Y. Jun Kang , S. Yang , Artif. Organs 2010 , 34 ( 11 ), 996 – 1002 ; b) E. Ozkumur , A. M. Shah , J. C. Ciciliano , B. L. Emmink , D. T. Miyamoto , E. Brachtel , M. Yu , P. I. Chen , B. Morgan , J. Trautwein , A. Kimura , S. Sengupta , S. L. Stott , N. M. Karabacak , T. A. Barber , J. R. Walsh , K. Smith , P. S. Spuhler , J. P. Sullivan , R. J. Lee , D. T. Ting , X. Luo , A. T. Shaw , A. Bardia , L. V. Sequist , D. N. Louis , S. Maheswaran , R. Kapur , D. A. Haber , M. Toner , Sci. Transl. Med. 2013 , 5 ( 179 ), 179ra47 ; c) K. K. Zeming , S. Ranjan , Y. Zhang , Nat. Commun. 2013 , 4 , 1625 .
[13] a) S. Bose , R. Singh , M. Hanewich-Hollatz , C. Shen , C. H. Lee , D. M. Dorfman , J. M. Karp , R. Karnik , Sci. Rep. 2013 , 3 , 2329 ; b) W. Chen , S. Weng , F. Zhang , S. Allen , X. Li , L. Bao , R. H. Lam , J. A. Macoska , S. D. Merajver , J. Fu , ACS Nano 2013 , 7 ( 1 ), 566 – 75 ; c) U. A. Gurkan , S. Tasoglu , D. Akkaynak , O. Avci , S. Unluisler , S. Canikyan , N. Maccallum , U. Demirci , Adv. Healthc. Mater. 2012 , 1 ( 5 ), 661 – 8 .
[14] J. Hardwick , ISBT Science Series 2008 , 3 ( 2 ), 148 – 176 . [15] A. Kreuger , O. Akerblom , C. F. Hogman , Vox Sang. 1975 , 29 ( 2 ),
81 – 9 . [16] a) J. L. Lozada , N. Caplanis , P. Proussaefs , J. Willardsen ,
G. Kammeyer , J. Oral Implantol. 2001 , 27 ( 1 ), 38 – 42 ; b) D. Pasqualetti , A. Ghirardini , M. Cristina Arista , S. Vaglio , A. Fakeri , A. A. Waldman , G. Girelli , Transfus. Apher. Sci. 2004 , 30 ( 1 ), 23 – 8 .
[17] E. C. Rossi , T. L. Simon , l. Wiley online, Rossi’s principles of transfusion medicine , Wiley-Blackwell , Chichester, UK; Hoboken, NJ , 2009 .
[18] A. V. Chernyshev , P. A. Tarasov , K. A. Semianov , V. M. Nekrasov , A. G. Hoekstra , V. P. Maltsev , J. Theor. Biol. 2008 , 251 ( 1 ), 93 – 107 .
[19] J. S. de Bono , H. I. Scher , R. B. Montgomery , C. Parker , M. C. Miller , H. Tissing , G. V. Doyle , L. W. Terstappen , K. J. Pienta , D. Raghavan , Clin. Cancer Res. 2008 , 14 ( 19 ), 6302 – 9 .
[20] W. A. Bonner , H. R. Hulett , R. G. Sweet , L. A. Herzenberg , Rev. Sci. Instrum. 1972 , 43 ( 3 ), 404 – 9 .
[21] S. Miltenyi , W. Muller , W. Weichel , A. Radbruch , Cytometry 1990 , 11 ( 2 ), 231 – 8 .
[22] a) W. J. Allard , J. Matera , M. C. Miller , M. Repollet , M. C. Connelly , C. Rao , A. G. Tibbe , J. W. Uhr , L. W. Terstappen , Clin. Cancer Res. 2004 , 10 ( 20 ), 6897 – 904 ; b) P. A. Liberti , C. G. Rao , L. W. M. M. Terstappen , J. Magn. Magn. Mater. 2001 , 225 ( 1–2 ), 301 – 307 .
[23] a) S. Maheswaran , L. V. Sequist , S. Nagrath , L. Ulkus , B. Brannigan , C. V. Collura , E. Inserra , S. Diederichs , A. J. Iafrate , D. W. Bell , S. Digumarthy , A. Muzikansky , D. Irimia , J. Settleman , R. G. Tompkins , T. J. Lynch , M. Toner , D. A. Haber , N. Engl. J. Med. 2008 , 359 ( 4 ), 366 – 77 ; b) D. T. Miyamoto , R. J. Lee , S. L. Stott , D. T. Ting , B. S. Wittner , M. Ulman , M. E. Smas , J. B. Lord , B. W. Brannigan , J. Trautwein , N. H. Bander , C. L. Wu , L. V. Sequist , M. R. Smith , S. Ramaswamy , M. Toner , S. Maheswaran , D. A. Haber , Cancer Discov. 2012 , 2 ( 11 ), 995 – 1003 ; c) M. Yu , A. Bardia , B. S. Wittner , S. L. Stott , M. E. Smas , D. T. Ting , S. J. Isakoff , J. C. Ciciliano , M. N. Wells , A. M. Shah , K. F. Concannon , M. C. Donaldson , L. V. Sequist , E. Brachtel , D. Sgroi , J. Baselga , S. Ramaswamy , M. Toner , D. A. Haber , S. Maheswaran , Science 2013 , 339 ( 6119 ), 580 – 4 ; d) M. Yu , D. T. Ting , S. L. Stott , B. S. Wittner , F. Ozsolak , S. Paul , J. C. Ciciliano , M. E. Smas , D. Winokur , A. J. Gilman , M. J. Ulman , K. Xega , G. Contino , B. Alagesan , B. W. Brannigan , P. M. Milos ,
Acknowledgements
We acknowledge valuable comments and suggestions on the manuscript from group members of the Integrated Biosystems and Biomechanics Laboratory. We thank Angela Hu, Mei Ki Cheung and Krystal Huijiao Guan for their assistance with the manuscript prep-aration. Work in Dr. Fu’s lab is supported by the National Science Foundation (CMMI 1129611, CBET 1149401, ECCS 1231826, CBET 1263889), the National Institute of Health (1R21HL114011), the American Heart Association (12SDG12180025), and the Depart-ment of Mechanical Engineering at the University of Michigan, Ann Arbor. Finally, we extend our apologies to colleagues in the fi eld whose work we were unable to discuss or cite formally because of space constraints and imposed reference limitations.
[1] A. J. Bruce Alberts , J. Lewis , M. Raff , K. Roberts , P. Walter , Table 22–1, Blood Cells . In Molecular Biology of the Cell , 4th ed. , New York : Garland Science , 2002 .
[2] a) P. Emery , T. Dorner , Ann. Rheum. Dis. 2011 , 70 ( 12 ), 2063 – 70 ; b) H. N. Iskandar , M. A. Ciorba , Transl. Res. 2012 , 159 ( 4 ), 313 – 25 ; c) J. M. Rhea , R. J. Molinaro , MLO. Med. Lab. Obs. 2011 , 43 ( 3 ), 10 – 2 , 16, 18; quiz 20, 22; d) P. Szodoray , Z. Szabo , A. Kapitany , A. Gyetvai , G. Lakos , S. Szanto , G. Szucs , Z. Szekanecz , Autoimmun. Rev. 2010 , 9 ( 3 ), 140 – 3 ; e) R. S. Vasan , Circulation 2006 , 113 ( 19 ), 2335 – 62 .
[3] a) W. F. Ganong , Review of medical physiology , Lange Medical Books/McGraw-Hill , New York 2003 ; b) M. L. Turgeon , Clinical Hematology: Theory and Procedures , Lippincott Williams & Wilkins , 2004 .
[4] a) Q. Zhou , T. Kwa , Y. Liu , A. Revzin , Expert. Rev. Anti. Infect. Ther. 2012 , 10 ( 10 ), 1079 – 81 ; b) W. Chen , N. T. Huang , X. Li , Z. T. Yu , K. Kurabayashi , J. Fu , Front. Oncol. 2013 , 3 , 98 .
[5] a) M. Cristofanilli , G. T. Budd , M. J. Ellis , A. Stopeck , J. Matera , M. C. Miller , J. M. Reuben , G. V. Doyle , W. J. Allard , L. W. Terstappen , D. F. Hayes , N. Engl. J. Med. 2004 , 351 ( 8 ), 781 – 91 ; b) M. C. Liu , P. G. Shields , R. D. Warren , P. Cohen , M. Wilkinson , Y. L. Ottaviano , S. B. Rao , J. Eng-Wong , F. Seillier-Moiseiwitsch , A. M. Noone , C. Isaacs , J. Clin. Oncol. 2009 , 27 ( 31 ), 5153 – 9 ; c) M. C. Miller , G. V. Doyle , L. W. Terstappen , J. Oncol. 2010 , 2010 , 617421 .
[6] M. L. Jones , J. Siddiqui , K. J. Pienta , R. H. Getzenberg , Prostate 2013 , 73 ( 2 ), 176 – 81 .
[7] a) A. M. Greenbaum , D. C. Link , Leukemia 2011 , 25 ( 2 ), 211 – 7 ; b) J. Thomas , F. Liu , D. C. Link , Curr. Opin. Hematol. 2002 , 9 ( 3 ), 183 – 9 .
[8] a) D. W. Bianchi , Br. J. Haematol. 1999 , 105 ( 3 ), 574 – 83 ; b) E. Guetta , M. J. Simchen , K. Mammon-Daviko , D. Gordon , A. Aviram-Goldring , N. Rauchbach , G. Barkai , Stem Cells Dev. 2004 , 13 ( 1 ), 93 – 9 ; c) S. S. Wachtel , L. P. Shulman , D. Sammons , Clin. Genet. 2001 , 59 ( 2 ), 74 – 9 .
[9] U. Dharmasiri , M. A. Witek , A. A. Adams , S. A. Soper , Annu. Rev. Anal. Chem. 2010 , 3 , 409 – 31 .
[10] a) J. Autebert , B. Coudert , F. C. Bidard , J. Y. Pierga , S. Descroix , L. Malaquin , J. L. Viovy , Methods 2012 , 57 ( 3 ), 297 – 307 ; b) A. A. S. Bhagat , H. Bow , H. W. Hou , S. J. Tan , J. Han , C. T. Lim , Med. Biol. Eng. Comput. 2010 , 48 ( 10 ), 999 – 1014 ; c) I. Cima , C. W. Yee , F. S. Iliescu , W. M. Phyo , K. H. Lim , C. Iliescu , M. H. Tan , Biomicrofl uidics 2013 , 7 ( 1 ); d) Y. Dong , A. M. Skelley , K. D. Merdek , K. M. Sprott , C. S. Jiang , W. E. Pierceall , J. Lin , M. Stocum , W. P. Carney , D. A. Smirnov , J. Mol. Diagn. 2013 ,
reviews[42] a) T. Songjaroen , W. Dungchai , O. Chailapakul , C. S. Henry ,
W. Laiwattanapaisal , Lab Chip 2012 , 12 ( 18 ), 3392 – 8 ; b) S. J. Vella , P. Beattie , R. Cademartiri , A. Laromaine , A. W. Martinez , S. T. Phillips , K. A. Mirica , G. M. Whitesides , Anal. Chem. 2012 , 84 ( 6 ), 2883 – 91 .
[43] T. F. Didar , K. Li , M. Tabrizian , T. Veres , Lab Chip 2013 , 13 ( 13 ), 2615 – 22 .
[44] M. S. Kim , T. S. Sim , Y. J. Kim , S. S. Kim , H. Jeong , J. M. Park , H. S. Moon , S. I. Kim , O. Gurel , S. S. Lee , J. G. Lee , J. C. Park , Lab Chip 2012 , 12 ( 16 ), 2874 – 80 .
[45] M. X. Lin , K. A. Hyun , H. S. Moon , T. S. Sim , J. G. Lee , J. C. Park , S. S. Lee , H. I. Jung , Biosensors Bioelectron. 2013 , 40 ( 1 ), 63 – 7 .
[46] D. Di Carlo , Lab Chip 2009 , 9 ( 21 ), 3038 – 46 . [47] D. Di Carlo , D. Irimia , R. G. Tompkins , M. Toner , Proc. Natl. Acad.
Sci. U. S. A. 2007 , 104 ( 48 ), 18892 – 7 . [48] A. A. Bhagat , H. W. Hou , L. D. Li , C. T. Lim , J. Han , Lab Chip 2011 ,
11 ( 11 ), 1870 – 8 . [49] H. W. Hou , M. E. Warkiani , B. L. Khoo , Z. R. Li , R. A. Soo ,
D. S. Tan , W. T. Lim , J. Han , A. A. Bhagat , C. T. Lim , Sci. Rep. 2013 , 3 , 1259 .
[50] L. Wu , G. Guan , H. W. Hou , A. A. Bhagat , J. Han , Anal. Chem. 2012 , 84 ( 21 ), 9324 – 31 .
[51] V. Parichehreh , K. Medepallai , K. Babbarwal , P. Sethu , Lab Chip 2013 , 13 ( 5 ), 892 – 900 .
[52] T. Tanaka , T. Ishikawa , K. Numayama-Tsuruta , Y. Imai , H. Ueno , N. Matsuki , T. Yamaguchi , Lab Chip 2012 , 12 ( 21 ), 4336 – 43 .
[53] a) T. A. Balbino , N. T. Aoki , A. A. M. Gasperini , C. L. P. Oliveira , A. R. Azzoni , L. P. Cavalcanti , L. G. de la Torre , Chem. Eng. J. 2013 , 226 , 423 – 433 ; b) S. Hou , S. Wang , Z. T. F. Yu , N. Q. M. Zhu , K. Liu , J. Sun , W. Y. Lin , C. K. F. Shen , X. Fang , H. R. Tseng , Angew. Chem. Int. Ed. Engl. 2008 , 47 ( 6 ), 1072 – 1075 ; c) S. Takayama , J. C. McDonald , E. Ostuni , M. N. Liang , P. J. A. Kenis , R. F. Ismagilov , G. M. Whitesides , Proc. Natl. Acad. Sci. U. S. A. 1999 , 96 ( 10 ), 5545 – 5548 ; d) Y. S. Torisawa , B. Mosadegh , G. D. Luker , M. Morell , K. S. O’Shea , S. Takayama , Integr. Biol. 2009 , 1 ( 11–12 ), 649 – 654 .
[54] a) P. Sethu , L. L. Moldawer , M. N. Mindrinos , P. O. Scumpia , C. L. Tannahill , J. Wilhelmy , P. A. Efron , B. H. Brownstein , R. G. Tompkins , M. Toner , Anal. Chem. 2006 , 78 ( 15 ), 5453 – 61 ; b) P. Sethu , M. Anahtar , L. L. Moldawer , R. G. Tompkins , M. Toner , Anal. Chem. 2004 , 76 ( 21 ), 6247 – 53 .
[55] a) S. Choi , T. Ku , S. Song , C. Choi , J. K. Park , Lab Chip 2011 , 11 ( 3 ), 413 – 8 ; b) S. Choi , S. Song , C. Choi , J. K. Park , Lab Chip 2007 , 7 ( 11 ), 1532 – 8 ; c) S. Choi , S. Song , C. Choi , J. K. Park , Small 2008 , 4 ( 5 ), 634 – 41 .
[56] J. A. Bernate , C. Liu , L. Lagae , K. Konstantopoulos , G. Drazer , Lab Chip 2013 , 13 ( 6 ), 1086 – 92 .
[57] J. A. Davis , D. W. Inglis , K. J. Morton , D. A. Lawrence , L. R. Huang , S. Y. Chou , J. C. Sturm , R. H. Austin , Proc. Natl. Acad. Sci. U. S. A. 2006 , 103 ( 40 ), 14779 – 84 .
[58] J. P. Beech , S. H. Holm , K. Adolfsson , J. O. Tegenfeldt , Lab Chip 2012 , 12 ( 6 ), 1048 – 51 .
[59] R. Huang , T. A. Barber , M. A. Schmidt , R. G. Tompkins , M. Toner , D. W. Bianchi , R. Kapur , W. L. Flejter , Prenat. Diagn. 2008 , 28 ( 10 ), 892 – 9 .
[60] S. H. Holm , J. P. Beech , M. P. Barrett , J. O. Tegenfeldt , Lab Chip 2011 , 11 ( 7 ), 1326 – 32 .
[61] K. Loutherback , J. D’Silva , L. Liu , A. Wu , R. H. Austin , J. C. Sturm , AIP Adv. 2012 , 2 ( 4 ), 4 2107 .
[62] R. Zhou , H. C. Chang , J. Colloid Interface Sci. 2005 , 287 ( 2 ), 647 – 56 .
[63] A. W. Browne , L. Ramasamy , T. P. Cripe , C. H. Ahn , Lab Chip 2011 , 11 ( 14 ), 2440 – 6 .
[64] M. Faivre , M. Abkarian , K. Bickraj , H. A. Stone , Biorheology 2006 , 43 ( 2 ), 147 – 59 .
[65] S. S. Shevkoplyas , T. Yoshida , L. L. Munn , M. W. Bitensky , Anal. Chem. 2005 , 77 ( 3 ), 933 – 7 .
D. P. Ryan , L. V. Sequist , N. Bardeesy , S. Ramaswamy , M. Toner , S. Maheswaran , D. A. Haber , Nature 2012 , 487 ( 7408 ), 510 – 3 .
[24] a) B. A. Frederick , B. A. Helfrich , C. D. Coldren , D. Zheng , D. Chan , P. A. Bunn Jr. , D. Raben , Mol. Cancer Ther. 2007 , 6 ( 6 ), 1683 – 91 ; b) M. Santisteban , J. M. Reiman , M. K. Asiedu , M. D. Behrens , A. Nassar , K. R. Kalli , P. Haluska , J. N. Ingle , L. C. Hartmann , M. H. Manjili , D. C. Radisky , S. Ferrone , K. L. Knutson , Cancer Res. 2009 , 69 ( 7 ), 2887 – 95 .
[25] B. T. van der Gun , L. J. Melchers , M. H. Ruiters , L. F. de Leij , P. M. McLaughlin , M. G. Rots , Carcinogenesis 2010 , 31 ( 11 ), 1913 – 21 .
[26] a) R. M. Bohmer , H. P. Stroh , K. L. Johnson , E. S. LeShane , D. W. Bianchi , Fetal Diagn. Ther. 2002 , 17 ( 2 ), 83 – 9 ; b) G. A. Challen , N. Boles , K. K. Lin , M. A. Goodell , Cytometry A 2009 , 75 ( 1 ), 14 – 24 ; c) M. Choolani , K. O’Donoghue , D. Talbert , S. Kumar , I. Roberts , E. Letsky , P. R. Bennett , N. M. Fisk , Mol. Hum. Reprod. 2003 , 9 ( 4 ), 227 – 35 ; d) A. Mavrou , E. Kouvidi , A. Antsaklis , A. Souka , S. Kitsiou Tzeli , A. Kolialexi , Prenat. Diagn. 2007 , 27 ( 2 ), 150 – 3 ; e) B. Prieto , M. Candenas , R. Venta , J. H. Ladenson , F. V. Alvarez , Clin. Chem. Lab. Med. 2002 , 40 ( 7 ), 667 – 72 .
[27] a) J. SooHoo , G. Walker , Conf. Proc. IEEE Eng. Med. Biol. Soc. 2007 , 2007 , 6319 – 22 ; b) J. R. Soohoo , G. M. Walker , Biomed. Microdevices 2009 , 11 ( 2 ), 323 – 9 .
[28] a) O. G. Helleso , P. Lovhaugen , A. Z. Subramanian , J. S. Wilkinson , B. S. Ahluwalia , Lab Chip 2012 , 12 ( 18 ), 3436 – 40 ; b) A. Kasukurti , M. Potcoava , S. A. Desai , C. Eggleton , D. W. M. Marr , Opt. Express 2011 , 19 ( 11 ), 10377 – 10386 ; c) K. Svoboda , S. M. Block , Annu. Rev. Biophys. Biomol. Struct. 1994 , 23 , 247 – 285 .
[29] X. B. Zhang , Z. Q. Wu , K. Wang , J. Zhu , J. J. Xu , X. H. Xia , H. Y. Chen , Anal. Chem. 2012 , 84 ( 8 ), 3780 – 6 .
[30] a) J. W. Song , S. P. Cavnar , A. C. Walker , K. E. Luker , M. Gupta , Y. C. Tung , G. D. Luker , S. Takayama , PLoS One 2009 , 4 ( 6 ), e5756 ; b) X. Zheng , L. S. Cheung , J. A. Schroeder , L. Jiang , Y. Zohar , Lab Chip 2011 , 11 ( 20 ), 3431 – 9 .
[31] a) D. R. Gossett , W. M. Weaver , A. J. Mach , S. C. Hur , H. T. Tse , W. Lee , H. Amini , D. Di Carlo , Anal. Bioanal. Chem. 2010 , 397 ( 8 ), 3249 – 67 ; b) M. Kersaudy-Kerhoas , E. Sollier , Lab Chip 2013 , 13 ( 17 ), 3323 – 46 .
[32] P. Wilding , J. Pfahler , H. H. Bau , J. N. Zemel , L. J. Kricka , Clin. Chem. 1994 , 40 ( 1 ), 43 – 7 .
[33] D. Lee , P. Sukumar , A. Mahyuddin , M. Choolani , G. Xu , J. Chro-matogr. A 2010 , 1217 ( 11 ), 1862 – 6 .
[34] P. Sethu , A. Sin , M. Toner , Lab Chip 2006 , 6 ( 1 ), 83 – 9 . [35] H. Mohamed , L. D. McCurdy , D. H. Szarowski , S. Duva ,
J. N. Turner , M. Caggana , IEEE Trans. NanoBiosci. 2004 , 3 ( 4 ), 251 – 6 .
[36] P. Preira , V. Grandne , J. M. Forel , S. Gabriele , M. Camara , O. Theodoly , Lab Chip 2013 , 13 ( 1 ), 161 – 70 .
[37] S. J. Tan , L. Yobas , G. Y. Lee , C. N. Ong , C. T. Lim , Biomed. Micro-devices 2009 , 11 ( 4 ), 883 – 92 .
[38] S. M. McFaul , B. K. Lin , H. Ma , Lab Chip 2012 , 12 ( 13 ), 2369 – 76 . [39] a) N. T. Huang , W. Chen , B. R. Oh , T. T. Cornell , T. P. Shanley ,
J. Fu , K. Kurabayashi , Lab Chip 2012 , 12 ( 20 ), 4093 – 101 ; b) W. Chen , N. T. Huang , X. Li , Z. T. F. Yu , K. Kurabayashi , J. Fu , Frontiers in Tumor Immunity (Cancer Immunotherapy & Immuno-monitoring: Mechanism, Treatment, Diagnosis, and Emerging Tools) 2003 , In press .
[40] a) T. Xu , B. Lu , Y. C. Tai , A. Goldkorn , Cancer Res. 2010 , 70 ( 16 ), 6420 – 6 ; b) H. K. Lin , S. Zheng , A. J. Williams , M. Balic , S. Groshen , H. I. Scher , M. Fleisher , W. Stadler , R. H. Datar , Y. C. Tai , R. J. Cote , Clin. Cancer Res. 2010 , 16 ( 20 ), 5011 – 8 .
[41] M. Hosokawa , M. Asami , S. Nakamura , T. Yoshino , N. Tsujimura , M. Takahashi , S. Nakasono , T. Tanaka , T. Matsunaga , Bio-technol. Bioeng. 2012 , 109 ( 8 ), 2017 – 24 .
Received: September 7, 2013 Revised: December 16, 2013
Published online: February 10, 2014
[66] S. Yang , A. Undar , J. D. Zahn , Lab Chip 2006 , 6 ( 7 ), 871 – 80 . [67] a) G. Kretzmer , K. Schugerl , Appl. Microbiol. Biotechnol. 1991 ,
34 ( 5 ), 613 – 6 ; b) Y. S. Li , J. H. Haga , S. Chien , J. Biomech. 2005 , 38 ( 10 ), 1949 – 71 .
[68] K. T. Kotz , W. Xiao , C. Miller-Graziano , W. J. Qian , A. Russom , E. A. Warner , L. L. Moldawer , A. De , P. E. Bankey , B. O. Petritis , D. G. Camp , 2nd , A. E. Rosenbach , J. Goverman , S. P. Fagan , B. H. Brownstein , D. Irimia , W. Xu , J. Wilhelmy , M. N. Mindrinos , R. D. Smith , R. W. Davis , R. G. Tompkins , M. Toner , Nat. Med. 2010 , 16 ( 9 ), 1042 – 7 .
[69] S. Nagrath , L. V. Sequist , S. Maheswaran , D. W. Bell , D. Irimia , L. Ulkus , M. R. Smith , E. L. Kwak , S. Digumarthy , A. Muzikansky , P. Ryan , U. J. Balis , R. G. Tompkins , D. A. Haber , M. Toner , Nature 2007 , 450 ( 7173 ), 1235 – 9 .
[70] S. L. Stott , C. H. Hsu , D. I. Tsukrov , M. Yu , D. T. Miyamoto , B. A. Waltman , S. M. Rothenberg , A. M. Shah , M. E. Smas , G. K. Korir , F. P. Floyd , A. J. Gilman , J. B. Lord , D. Winokur , S. Springer , D. Irimia , S. Nagrath , L. V. Sequist , R. J. Lee , K. J. Isselbacher , S. Maheswaran , D. A. Haber , M. Toner , Proc. Natl. Acad. Sci. U. S. A. 2010 , 107 ( 43 ), 18392 – 18397 .
[71] S. Wang , K. Liu , J. Liu , Z. T. Yu , X. Xu , L. Zhao , T. Lee , E. K. Lee , J. Reiss , Y. K. Lee , L. W. Chung , J. Huang , M. Rettig , D. Seligson , K. N. Duraiswamy , C. K. Shen , H. R. Tseng , Angew. Chem. Int. Ed. Engl. 2011 , 50 ( 13 ), 3084 – 8 .
[72] A. D. Hughes , J. Mattison , J. D. Powderly , B. T. Greene , M. R. King , J. Vis. Exp. 2012 , ( 64 ), e4248 .
[73] W. Sheng , T. Chen , R. Kamath , X. Xiong , W. Tan , Z. H. Fan , Anal. Chem. 2012 , 84 ( 9 ), 4199 – 206 .
[74] S. Choi , J. M. Karp , R. Karnik , Lab Chip 2012 , 12 ( 8 ), 1427 – 30 . [75] C. Launiere , M. Gaskill , G. Czaplewski , J. H. Myung , S. Hong ,
D. T. Eddington , Anal. Chem. 2012 , 84 ( 9 ), 4022 – 8 . [76] P. Li , Y. Gao , D. Pappas , Anal. Chem. 2012 , 84 ( 19 ), 8140 – 8 . [77] a) J. L. Johnson , M. B. Jones , S. O. Ryan , B. A. Cobb , Trends
Immunol. 2013 , 34 ( 6 ), 290 – 8 ; b) A. Varki , J. D. Esko , K. J. Colley , Cellular Organization of Glycosylation . In Essentials of Glyco-biology , 2nd ed. , Cold Spring Harbor (NY) : Cold Spring Harbor Laboratory Press , 2009 .
[78] a) D. J. Laderach , L. Gentilini , F. M. Jaworski , D. Compagno , Prostate Cancer 2013 , 2013 , 519436 ; b) V. Padler-Karavani , Cancer Lett. 2013 .
[79] a) G. Simone , N. Malara , V. Trunzo , G. Perozziello , P. Neuzil , M. Francardi , L. Roveda , M. Renne , U. Prati , V. Mollace , A. Manz , E. Di Fabrizio , Small 2013 , 9 ( 12 ), 2152 – 2161 ; b) G. Simone , P. Neuzil , G. Perozziello , M. Francardi , N. Malara , E. Di Fabrizio , A. Manz , Lab Chip 2012 , 12 ( 8 ), 1500 – 1507 .
[80] H. Zhu , G. Stybayeva , M. Macal , E. Ramanculov , M. D. George , S. Dandekar , A. Revzin , Lab Chip 2008 , 8 ( 12 ), 2197 – 205 .
[81] U. A. Gurkan , T. Anand , H. Tas , D. Elkan , A. Akay , H. O. Keles , U. Demirci , Lab Chip 2011 , 11 ( 23 ), 3979 – 89 .
[82] S. Hou , L. Zhao , Q. Shen , J. Yu , C. Ng , X. Kong , D. Wu , M. Song , X. Shi , X. Xu , W. H. OuYang , R. He , X. Z. Zhao , T. Lee , F. C. Brunicardi , M. A. Garcia , A. Ribas , R. S. Lo , H. R. Tseng , Angew. Chem. Int. Ed. Engl. 2013 , 52 ( 12 ), 3379 – 83 .
[83] S. Ariyasu , K. Hanaya , E. Watanabe , T. Suzuki , K. Horie , M. Hayase , R. Abe , S. Aoki , Langmuir 2012 , 28 ( 36 ), 13118 – 26 .
[84] a) J. H. Kang , S. Krause , H. Tobin , A. Mammoto , M. Kanapathipillai , D. E. Ingber , Lab Chip 2012 , 12 ( 12 ), 2175 – 81 ; b) Y. Y. Huang , K. Hoshino , P. Chen , C. H. Wu , N. Lane , M. Huebschman , H. Liu , K. Sokolov , J. W. Uhr , E. P. Frenkel , J. X. Zhang , Biomed. Microdevices 2013 , 15 ( 4 ), 673 – 81 ; c) B. P. Casavant , L. N. Strotman , J. J. Tokar , S. M. Thiede , A. M. Traynor , J. S. Ferguson , J. M. Lang , D. J. Beebe , Lab Chip 2013 ; d) B. P. Casavant , D. J. Guckenberger , S. M. Berry , J. T. Tokar , J. M. Lang , D. J. Beebe , Lab Chip 2013 , 13 ( 3 ), 391 – 6 .
[85] D. Issadore , H. Shao , J. Chung , A. Newton , M. Pittet , R. Weissleder , H. Lee , Lab Chip 2011 , 11 ( 1 ), 147 – 51 .
[86] A. E. Saliba , L. Saias , E. Psychari , N. Minc , D. Simon , F. C. Bidard , C. Mathiot , J. Y. Pierga , V. Fraisier , J. Salamero , V. Saada , F. Farace , P. Vielh , L. Malaquin , J. L. Viovy , Proc. Natl. Acad. Sci. U. S. A. 2010 , 107 ( 33 ), 14524 – 9 .
[87] K. H. Han , A. B. Frazier , IEE Proc. Nanobiotechnol. 2006 , 153 ( 4 ), 67 – 73 .
[88] J. Nam , H. Huang , H. Lim , C. Lim , S. Shin , Anal. Chem. 2013 , 85 ( 15 ), 7316 – 23 .
[89] D. R. Arifi n , L. Y. Yeo , J. R. Friend , Biomicrofl uidics 2007 , 1 ( 1 ), 14103 .
[90] Y. J. Kang , D. Q. Li , S. A. Kalams , J. E. Eid , Biomed. Microdevices 2008 , 10 ( 2 ), 243 – 249 .
[91] G. J. Shah , J. L. Veale , Y. Korin , E. F. Reed , H. A. Gritsch , C. J. Kim , Biomicrofl uidics 2010 , 4 ( 4 ).
[92] a) J. Gao , X. F. Yin , Z. L. Fang , Lab Chip 2004 , 4 ( 1 ), 47 – 52 ; b) X. Xuan , D. Li , Electrophoresis 2005 , 26 ( 18 ), 3552 – 60 .
[93] M. W. Wang , Electrophoresis 2012 , 33 ( 5 ), 780 – 7 . [94] T. Z. Jubery , P. Dutta , Electrophoresis 2013 , 34 ( 5 ), 643 – 50 . [95] J. Gao , R. Riahi , M. L. Sin , S. Zhang , P. K. Wong , Analyst 2012 ,
137 ( 22 ), 5215 – 21 . [96] M. D. Vahey , J. Voldman , Anal. Chem. 2009 , 81 ( 7 ), 2446 –
2455 . [97] P. Augustsson , C. Magnusson , M. Nordin , H. Lilja , T. Laurell ,
Anal. Chem. 2012 , 84 ( 18 ), 7954 – 62 . [98] J. Dykes , A. Lenshof , I. B. Astrand-Grundstrom , T. Laurell ,
S. Scheding , PLoS One 2011 , 6 ( 8 ), e23074 . [99] A. Lenshof , A. Ahmad-Tajudin , K. Jaras , A. M. Sward-Nilsson ,
L. Aberg , G. Marko-Varga , J. Malm , H. Lilja , T. Laurell , Anal. Chem. 2009 , 81 ( 15 ), 6030 – 7 .
[100] M. Nordin , T. Laurell , Lab Chip 2012 , 12 ( 22 ), 4610 – 6 . [101] M. A. Burguillos , C. Magnusson , M. Nordin , A. Lenshof ,
P. Augustsson , M. J. Hansson , E. Elmer , H. Lilja , P. Brundin , T. Laurell, T. Deierborg, PLoS One 2013, 8(5), e64233.